PROJECT SUMMARY - Overview Fusion oncoproteins (FO) arising from the rearrangement of Nucleoporin 98 (NUP98) to a diverse range of partner genes are a hallmark of multiple subtypes of high risk myeloid malignancies in children and adolescents, including acute erythroleukemia and acute megakaryoblastic leukemia. Such leukemias are associated with poor outcome, and more effective therapies are required. The long-term goal of this Center for Experimental and Preclinical Modeling of NUP98 Leukemia is to determine the molecular role, and potential vulnerabilities, of NUP98-fusion oncoproteins in leukemia through the development and interrogation of human and mouse model systems, and to develop and test novel therapeutic approaches. The Center has assembled a consortium of investigators with complementary expertise in genomic characterization, leukemia modeling, chromatin biology, structural biology and preclinical modeling. The Center is led by Dr Charles Mullighan of St Jude Children?s Research Hospital together with Project Co-Leaders Drs Scott Armstrong (Dana Farber Cancer Institute), Taosheng Chen (St Jude), Alex Kentsis (Memorial Sloan Kettering Cancer Institute), Jeffery Klco (St Jude) and Richard Kriwacki (St Jude). These investigators will co-Lead four projects performing experimental modeling (Project 1), investigating chromatin biology (Project 2), phase separation (Project 3) and drug development (Project 4) that will address the following questions and unmet needs: (1) to define how NUP98 fusions drive leukemogenesis using complementary, integrative engineered models and proteomic, transcriptomic and epigenomic profiles; (2) to develop genetically faithful engineered mouse, human and xenograft models of NUP98 leukemia for mechanistic interrogation and preclinical modeling; (3) to define the macromolecular chromatin and protein complexes assembled by NUP98 FOs; (4) to define the role of phase separation in NUP98 FO leukemogenesis; (5) to define the dependencies and vulnerabilities of NUP98-rearranged leukemic cells; and (6) to develop more effective therapeutic strategies for NUP98-rearranged leukemia. The Center is supported by an Administrative Core A that with an External Advisory Board will oversee project progress interaction and reporting, a Genome Editing Core B led by Dr David Chen (City of Hope) that provides expertise and support for CRISPR/Cas9 genome editing scans and screens; and a Chemistry Core C led by Dr Zoran Rankovic (St Jude) that supports chemical library screen and drug design and development. This highly integrated, interactive and synergistic Center will provide fundamental insights into the mechanisms of oncogenesis of the different classes of NUP98 FO, and new therapeutic modalities that will be validated in preclinical models and translated into human clinical trials. All data and resources will be made freely available by websites, data deposition, and establishment of resource and genomic data portals to the FusOnC2 consortium and broad scientific community.